Abstract:
:The purpose of this study was to explore the potential of using cationic polyethylenimine (PEI) to deliver green fluorescent protein (GFP) to protozoan parasite Toxoplasma gondii. PEI/DNA polyplexes were formed using branched PEI and pEGFP-N1 plasmid with various N/P ratios that ranged from 5 to 50. With the increment of N/P ratio, the average size of formed PEI/DNA polyplexes determined by dynamic light scattering analysis decreased from 306 to 203 nm, while the surface charge of polyplexes obtained by zeta potential measurements increased from 20.2 to 36.7 mV. Gene transfection efficiency modulated by N/P ratio was determined, indicating PEI/DNA polyplexes were capable of transfecting parasites. The maximal GFP expression was observed 8 h post-transfection using N/P ratio of 30. To demonstrate the infectivity and potential use of GFP-expressing T. gondii, transfected parasites were inoculated to the monolayer of human foreskin fibroblast (HFF) cells. GFP-expressing tachyzoites were observed in intracellular milieu of the infected HFF cells one day after the infection. After 12-day culture, the bradyzoites expressing GFP within cysts were clearly visualized extracellularly. Our results revealed that PEI can be harnessed as an effective and inexpensive reagent to construct GFP-expressing T. gondii which has potential uses such as the study of interconversion stages and antimicrobial drug screening.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Salehi N,Peng CAdoi
10.1016/j.mimet.2012.08.001subject
Has Abstractpub_date
2012-10-01 00:00:00pages
133-7issue
1eissn
0167-7012issn
1872-8359pii
S0167-7012(12)00260-6journal_volume
91pub_type
杂志文章abstract::A rapid, simple and sensitive spectrofluorimetric technique was developed for monitoring total lipids in hyphae of the riboflavin-overproducing fungus Eremothecium ashbyii using the fluorescent probe Nile blue in an aqueous system, avoiding the interference due to autofluorescent riboflavin. The existing methodologies...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/s0167-7012(03)00137-4
更新日期:2003-10-01 00:00:00
abstract::Polyhydroxyalkanoates (PHAs) are important for biodegradable plastic production, and prokaryotes play a very important role in PHA production. PHA synthase is a key enzyme for the polymerization of PHAs. There are four classes of PHA synthase. The phaC gene is necessary for the production of all classes of PHA synthas...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2015.06.012
更新日期:2015-09-01 00:00:00
abstract::A protein database was constructed and validated with identification rate over 90% for the 4 most common Gram-negative bacteria on agar plates. By protein masses comparison, 120 bacteria of the 4 species from blood culture bottles were identified. The concordance was high (Kappa=0.906) between our method and conventio...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2014.07.021
更新日期:2014-10-01 00:00:00
abstract::Bacteriocin activity can be quantified by a simple parallel line model based on an agar diffusion assay. The quotient between the inhibition zone area and the sensitivity of the indicator bacterium can be used to compare different bacteriocins. Deferred calculations of relative potencies are possible, once purified co...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2005.01.006
更新日期:2005-07-01 00:00:00
abstract::A new concept for multiplex detection and quantification of microbes is here demonstrated on a range of infectious fungal species. Padlock probe methodology in conjunction with qPCR and Luminex technology was used for simultaneous detection of ten fungal species in one single experiment. By combining the multiplexing ...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2009.05.016
更新日期:2009-08-01 00:00:00
abstract::Immunoproteomic technology offers an exceptional tool to fill the blanks that still exist in diagnosis of Toxoplasma gondii infection despite its annotated sequence. The pitfalls of serological assays and current immunoproteomic approaches are accentuated, and new approaches are presented to improve the signal and to ...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2015.09.011
更新日期:2015-12-01 00:00:00
abstract::A simple and sensitive method based on the use of colloidal chitin stained with Remazol Brilliant Blue R (RBB) is proposed to evaluate chitinase activity. If this colloidal-stained substrate is included as a carbon source in a liquid medium, this technique allows the selection or the comparison of chitinolytic microor...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2003.10.011
更新日期:2004-02-01 00:00:00
abstract::In wastewater, biological phosphate removal can fail because of the presence of glycogen accumulating organism (GAO), therefore measuring glycogen stored in microbial cultures provides information on the bacterial population type. Once glycogen is hydrolysed to glucose it was accurately measured using a human glucose ...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2014.03.002
更新日期:2014-05-01 00:00:00
abstract::The biotechnologically important white-rot fungus Physisporinus vitreus was co-cultivated with Agrobacterium tumefaciens AGL-1 carrying plasmids with nourseothricin resistance as the selectable marker gene and red fluorescence protein as a visual marker. Mitotically stable transformed isolates were obtained showing re...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2013.09.001
更新日期:2013-11-01 00:00:00
abstract::The purpose of this study was to develop a biosensor based on surface plasmon resonance (SPR) for the rapid identification of C. jejuni in broiler samples. We examined the specificity and sensitivity of commercial antibodies against C. jejuni with six Campylobacter strains and six non-Campylobacter bacterial strains. ...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2006.12.002
更新日期:2007-04-01 00:00:00
abstract::The antimicrobial activity of lime oil was significantly enhanced against Staphylococcus epidermidis in a pulsed electromagnetic field. These findings support other workers who found that electromagnetic fields augment antimicrobial activity. ...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2010.09.015
更新日期:2010-11-01 00:00:00
abstract::Modified linker-PCR primers were developed to enable complete sequencing of a DGGE band in one reaction. Commonly used bacterial and archaeal 16S rRNA gene PCR-DGGE primers were modified to contain linkers and sequencing primers. This protocol does not involve additional stages, and improves retrieval of sequence from...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2008.08.006
更新日期:2008-12-01 00:00:00
abstract::An effective microbial preservation technology for the long-term storage of viable prokaryotic cells is described. The method combines an almost instantaneous drying step with minimal stress to the cells during drying to maximize survival on rehydration. This is achieved by contact of a microlitre aliquot of a bacteri...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2005.02.014
更新日期:2005-10-01 00:00:00
abstract::The Carba NP test was developed to detect carbapenemase-producing Enterobacteriaceae, and uses imipenem as the reaction substrate. In Japan, IMP-6 metallo-β-lactamase (MBL) producers, which are usually resistant to meropenem but susceptible to imipenem, and IMP-1 MBL producers, which are usually resistant to both carb...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2016.12.015
更新日期:2017-02-01 00:00:00
abstract::Culture in enriched broth, as well as on a solid medium, is recommended for primary isolation of mycobacteria. With the introduction of liquid mycobacterial culture methods, a substantial workload regarding the identification of culture-recovered mycobacterial species, particularly Mycobacterium tuberculosis complex (...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2015.10.011
更新日期:2015-12-01 00:00:00
abstract::Contamination of cereals with Fusarium species is one of the major sources of mycotoxins in food and feed. Quantification of biomass of Fusarium species is essential to understand the interactions of individual species in disease development. In this study quantitative real-time PCR assays based on the elongation fact...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2008.10.016
更新日期:2009-03-01 00:00:00
abstract::Culture-independent studies using next generation sequencing have revolutionized microbial ecology, however, oomycete ecology in soils is severely lagging behind. The aim of this study was to improve and validate standard techniques for using high throughput sequencing as a tool for studying oomycete communities. The ...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2015.01.013
更新日期:2015-03-01 00:00:00
abstract::Race 3, biovar 2 strains of Ralstonia solanacearum are quarantined pathogens in Europe and Canada and Select Agent pathogens in the United States. The biovar classification of R. solanacearum strains is based on their biochemical abilities to utilize a carbohydrate panel. The standard biovar test uses bromothymol blue...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2011.12.007
更新日期:2012-02-01 00:00:00
abstract::Nanoparticles synthesized through the green route deserve special mention because this green technology is not only energy-efficient and cost-effective but also amenable to the environment. Various biological resources have been used for the generation of these 'green nanoparticles'. Biological wastes have also been f...
journal_title:Journal of microbiological methods
pub_type: 杂志文章,评审
doi:10.1016/j.mimet.2019.105766
更新日期:2019-12-01 00:00:00
abstract::The bclA gene codes for the protein backbone of the exosporium glycoprotein BclA of B. anthracis. BclA has a central collagen-like region formed by polymorphic GXX repeats and conserved amino- and carboxy-termini. It is noted here that the bclA gene is also present in the genome of Bacillus cereus and Bacillus thuring...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2005.04.032
更新日期:2006-01-01 00:00:00
abstract::The susceptibility protocol established in this study takes into account that complex media are capable to buffer H2O2 which otherwise may adulterate test results. Furthermore, we demonstrate that clinical isolates of Staphylococcus aureus showed a higher resistance against H2O2 than Streptococcus pyogenes after 30 mi...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2018.11.001
更新日期:2018-12-01 00:00:00
abstract::Effective biolistic transformation of intact conidia from the filamentous fungus Trichoderma reesei was achieved using the Bio-Rad Hepta Adaptor system with seven barrels for particle launch. Transformation frequencies of up to 39 colonies per microg of circular DNA and 37 colonies per microg of linear DNA were obtain...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/s0167-7012(02)00126-4
更新日期:2002-11-01 00:00:00
abstract::We monitored Chlamydia trachomatis growth in HeLa cells cultured with either DMEM or RPMI medium containing 10% FCS under 2% or 21% O2 conditions for 2 days. Bacterial numbers, host cell numbers, and fibrosis-related gene expression in the host cells were estimated by an inclusion forming unit assay, a cell counting a...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2018.09.007
更新日期:2018-10-01 00:00:00
abstract::Global public health is threatened by the emergence of potentially dangerous antibiotic drug-resistant strains of Mycobacterium tuberculosis. Point mutations in certain M. tuberculosis genes are associated with the resistance of M. tuberculosis strains to antibiotic drugs. The purpose of this study was to develop a su...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2005.04.026
更新日期:2005-12-01 00:00:00
abstract::The development of biofilms in static plates was monitored. Glass coupons were placed on agar covered with filter paper, which was inoculated with suspended bacteria. The viable cell density, biofilms matrix and biomass were quantified. The method is excellent for adhesion and material studies, due to its simplicity a...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2014.10.008
更新日期:2014-12-01 00:00:00
abstract::Culture-independent molecular techniques and advances in next generation sequencing (NGS) technologies make large-scale epidemiological studies on microbiota feasible. A challenge using NGS is to obtain high reproducibility and repeatability, which is mostly attained through robust amplification. We aimed to assess th...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2018.03.003
更新日期:2018-04-01 00:00:00
abstract::Our understanding of the molecular mechanisms of bacterial infection and pathogenesis are disproportionally derived from a small number of well-characterised species and strains. One reason for this is the enormous time and resources required to develop a new organism into experimental system that can be interrogated ...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2016.08.022
更新日期:2016-11-01 00:00:00
abstract::The limited availability of biological samples hinders phylogenetic efforts to define structural differences among various biological groups. A novel workflow enabling the analysis of protists in low cell numbers by electron microscopy (EM) is described with cysts of Giardia intestinalis, a single-celled eukaryotic pa...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2020.105888
更新日期:2020-05-01 00:00:00
abstract::We have developed a software tool, GenomeComp, for summarizing, parsing and visualizing the genome sequences comparison results derived from voluminous BLAST textual output. With GenomeComp, the variation between genomes can be easily highlighted, such as repeat regions, insertions, deletions and rearrangements of gen...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/s0167-7012(03)00094-0
更新日期:2003-09-01 00:00:00
abstract::Several studies have been conducted to develop rapid methods for quantification of lipid content in microalgae, as an alternative for time consuming gravimetric methods. Different studies showed that lipid staining with Nile Red in whole cell suspensions and subsequently quantification by the use of a spectrofluoromet...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2015.09.009
更新日期:2015-11-01 00:00:00